Assay Principle
The target antibody is coated on a 96-well microplate, creating a solid-phase carrier. Standard samples or test samples are added to the wells, where target antibodies bound to the solid-phase carrier’s antibody. Subsequently, microbial target antibodies are added, and after washing to remove any unbound biotinylated antibodies, HRP-labeled and avidin are added. After thorough washing, a TMB substrate is added for color development. TMB is converted into a blue color in the presence of peroxidase and ultimately into yellow under the influence of acid. The intensity of the color is directly proportional to the amount of target in the sample. The absorbance (O.D. value) is measured using an ELISA reader at a wavelength of 450 nm, and the sample concentration is calculated based on this measurement.
